Studies On Cell And Biochemistry At Gametophytes Stage Of Ovule In Pinus Tabulaeformis Carr.

The material for studies on molecular mechanism of genetic regulation in the course of ovule development and gametophytes forming were mainly from the finding and inducement of mutation. In the seed orchard of Pinus tabulaeformis Carr. in Xingcheng, Liaoning Province, the ovules of the clone 28 aborted because the mitosis failure of free nuclei in female gametophyte. Because of the stable genetic characteristic, the ovule were nice material for research of the mechanism of ovule developmental regulation. The research was a part of work from nation natural science sustentation fund for (30371144) and (30671655). Useing ovule of female sterile clone 28 of P. tabulaeformis as material and fertile clone 14 as contrast, the changes of soluble sugar, free amino acid and soluble protein's contents in two lines ovule and suspending culture cell which induced from ovule that treated with nitrogen, phosphor, calcium, sucrose, NAA and 6-BA were studied by the methods of phenol, ninhydrin and coomassie brilliant blue G-250 etc. And the effect of nitrogen, phosphor, calcium, sucrose, NAA and 6-BA on the cell division and protein expression were also studied. The differential proteins were searched by 2-dimensional electrophoresis database. The results were as follows:According material contents' change of ovule, saving and obtaining nutrient material and phytohormone of sterile line's ovule developmemt needed were less and not corresponding which made metabolize foul-up of carbohydrate , amino acid and protein, so leaded mechanism of senescence that falling of soluble sugar contents, accumulating of free amino acid and decomposing of soluble protein,import and export of material that ovule develops need were broken, forming of bio-molecule and mitosis of free nuclei gametophytes were effected, thus it can be seen that ovule development was baffled and abortion was happened. The change of soluble sugar, free amino acid and soluble proteins contents were looked as criterion of physiology and biochemistry on studying development mechanism of ovule for this experiment.The change of soluble sugar, free amino acid and soluble protein's contents of two clones cell were effected by nitrogen, phosphor, calcium, sucrose, NAA and 6-BA had relatively genic stabilization. The nutrition functionary of restraining cell division sort by size was nitrogen, sucrose, phosphor and calcium, and phytohormone was 6-BA and NAA. 25.4, 36.8, 36.9 and 42.2 KDa proteins might be related to free nuclei mitosis of sterility ovule, and 56.6 and 90.7KDa proteins might be related to cell cycle regulation.The search results of two clones ovular differential proteins showed that some of factors that peroxidation and bio-molecule damnification hurt frame and function of cell and producing ethane because sterile clone ovule was devoid of protected enzyme system. but GAs was fall, not having perfect glycolysis and hepatin abnormality producing made intermediate material less and energy shortage and lacking Ser/Thr kinase and Ca~+/CaM signal transduction system,having cysteine proteinase which urge programmed cell death, all of these made ovule development was baffled and abortion was happened.These results supplied reference for more studies on physiology and biochemistry of ovule development in P. tabulaeformis, and supplied material for studying the reason of free nuclei gametophytes' stop division on cell level.